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Restriction Enzymes Resource

Table 3.1 Relative Activity of Restriction Enzymes in Promega's 10X Buffers

The 10X Reaction Buffer supplied with each restriction enzyme is optimized to give 100% activity. In many cases, good activity is also obtained using one of Promega's 4-CORE® 10X Buffers (Cat.# R9921). Table 3.1 may be used to select the best buffer for digestions with multiple restriction enzymes. Enzyme activity is expressed as a percent of the activity obtained with the optimized buffer for each enzyme.

Promega
Enzyme 		Buffer Supplied with Enzyme 4 CORE® Buffers MULTI-
CORE™
Buffer		 Enzyme Assay
Temp.
A B C D
Aat II J 50-75% 10-25% <10% <10% <10% 37°C
Acc I G 50-75% 25-50% 25-50% 10-25% 25-50% 37°C
Acc III F <10% 10-25% 25-50% 25-50% <10% 65°C
Acc65 I D 10-25% 50-75% 75-100% 100% 100% 37°C
AccB7 I E 10-25% 50-75% 100%** <10% 100% 37°C
Age I K 25-50% 25-50% 25-50% 50-75% 100% 37°C
Alu I B 75-100% 100% 75-100% 10-25% 10-25% 37°C
Alw26 I C 10-25% 25-50% 100% 10-25% 75-100% 37°C
Alw44 I C <10% 25-50% 100% 25-50% 100% 37°C
Apa I A 100% 50-75% 50-75% <10% 75-100% 37°C
Ava I B 10-25% 100% 50-75% 25-50% <10% 37°C
Ava II C 50-75% 50-75% 100% 25-50% 25-50% 37°C
Bal I G 10-25% <10% <10% <10% <10% 37°C
BamH I E 75-100% ** 75-100% 75-100% 50-75% 75-100% 37°C
Ban I G 25-50% 25-50% 10-25% <10% 100% 50°C
Ban II E 75-100% 75-100% 75-100% 25-50% 100% 37°C
Bbu I A 100% 75-100% 75-100% <10% 100% 37°C
Bcl I C 10-25% 75-100% 100% 50-75% 10-25% 50°C
Bgl I D 10-25% 25-50% 75-100% 100% 100% 37°C
Bgl II D 25-50% 75-100% 75-100% 100% <10% 37°C
BsaM I D 10-25% 25-50% 50-75% 100% 25-50% 65°C
BsaO I C 10-25% 50-75% 100% 25-50% 100% 50°C
Bsp I286 I A 100% 50-75% 25-50% 10-25% 75-100% 37°C
BsrBR I H 10-25% 50-75%** 100%** 50-75% 100% 65°C
BsrS I D 10-25% 25-50% 10-25% 100% 100% 65°C

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Promega
Enzyme 		Buffer Supplied
with Enzyme 		  4 CORE® Buffers MULTI-
CORE™ Buffer		 Enzyme Assay
Temperature
A B C D
BssH II H 75-100% 50-75% 75-100% 50-75% 75-100% 50°C
Bst71 I D 10-25% 25-50% 25-50% 100% 10-25% 50°C
Bst98 I D <10% 10-25% 10-25% 100% 25-50% 37°C
BstE II D 25-50% 50-75% 50-75% 100% 100% 60°C
BstO I C 10-25% 25-50% 100% 25-50% <10% 60°C
BstX I D <10% 10-25% 25-50% 100% 10-25% 50°C
BstZ I D <10% <10% 10-25% 100% 10-25% 50°C
Bsu36 I E <10% 25-50% 50-75% 25-50% 50-75% 37°C
Cfo I B 75-100% 100% 75-100% 25-50% 100% 37°C
Cla I C 75-100% 75-100% 100% 75-100% 100% 37°C
Csp I K <10% 10-25% 25-50% 50-75% 10-25% 30°C
Csp45 I B 25-50% 100% 50-75% 25-50% 50-75% 37°C
Dde I D 25-50% 25-50% 50-75% 100% 25-50% 37°C
Dpn I B 50-75% 100% 75-100% 50-75% 100% 37°C
Dra I B 75-100% 100% 75-100% 50-75% 25-50% 37°C
EclHK I E <10% <10% 75-100% 10-25% 50-75% 37°C
Eco47 III D <10% 25-50% 50-75% 100% 25-50% 37°C
Eco52 I L <10% <10% 10-25% 25-50% <10% 37°C
EcoICR I B 10-25% 100% 75-100% <10% 100% 37°C
EcoR I H 25-50% 50-75% 50-75% 50-75% 100% 37°C
EcoR V D 10-25% 25-50% 50-75% 100% 100% 37°C
Fok I B 75-100% 100% 75-100% 25-50% 50-75% 37°C
Hae II B 50-75% 100% 50-75% 10-25% 100% 37°C
Hae III C 75-100% 75-100% 100% 50-75% 100% 37°C

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Promega
Enzyme 		Buffer Supplied
with Enzyme 		4 CORE® Buffers MULTI-
CORE™ Buffer		 Enzyme Assay
Temperature
A B C D
Hha I C 50-75% 75-100% 100% 50-75% 75-100% 37°C
Hinc II B 25-50% 100% 25-50% 50-75% 100% 37°C
Hind III E 25-50% 100% 75-100% 10-25% 50-75% 37°C
Hinf I B 50-75% 100% 75-100% 75-100% 50-75% 37°C
Hpa I J 25-50% 50-75% 25-50% 10-25% 100% 37°C
Hpa II A 100% 50-75% 50-75% 10-25% 100% 37°C
Hsp92 I F 10-25% 75-100% 50-75% 25-50% 10-25% 37°C
Hsp92 II K 10-25% 25-50% 25-50% <10% <10% 37°C
I-Ppo I I-Ppo I 10-25% 25-50% 25-50% 25-50% - 37°C
Kpn I J 100%** 25-50% 25-50% <10% 75-100% 37°C
Mbo I C 10-25% 75-100% 100% 50-75% <10% 37°C
Mbo II B 10-25% 100% 50-75% 75-100% 100% 37°C
Mlu I D 10-25% 25-50% 50-75% 100% 10-25% 37°C
Msp I B 75-100% 100% 75-100% 25-50% 25-50% 37°C
MspA1 I C 25-50% 100%** 100% 10-25% 100% 37°C
Nae I A 100% 50-75% 25-50% <10% 50-75% 37°C
Nar I G 75-100% 50-75% 75-100% 25-50% 50-75% 37°C
Nci I B 100%** 100% 25-50% 25-50% 50-75% 37°C
Nco I D 50-75% 75-100% 75-100% 100% 75-100% 37°C
Nde I D <10% <10% 25-50% 100% 25-50% 37°C
Nde II D <10% <10% 10-25% 100% 25-50% 37°C
NgoM IV MULTI-
CORE™		 100%** 100%** 100%** <10% 100% 37°C
Nhe I B 75-100% 100% 75-100% 10-25% 100% 37°C
Not I D <10% 10-25% 25-50% 100% 25-50% 37°C

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Promega
Enzyme 		Buffer Supplied
with Enzyme 		4 CORE® Buffers MULTI-
CORE™ Buffer 		Enzyme Assay
Temperature
A B C D
Nru I K <10% <10% <10% 50-75% 10-25% 37°C
Nsi I D 10-25% 50-75% 50-75% 100% 10-25% 37°C
Pst I H 10-25% 50-75% 50-75% 50-75% 25-50% 37°C
Pvu I D 10-25% 25-50% 50-75% 100% <10% 37°C
Pvu II B 25-50% 100% 50-75% 25-50% 50-75% 37°C
Rsa I C 75-100% 75-100% 100% <10% <10% 37°C
Sac I J 75-100% 25-50% 25-50% <10% 100% 37°C
Sac II C 100%* 50-75% 100% 50-75% <10% 37°C
Sal I D <10% 10-25% 25-50% 100% <10% 37°C
Sau3A I B 25-50% 100% 75-100% <10% 100% 37°C
Sau96 I C 25-50% 25-50% 100% 50-75% 50-75% 37°C
Sca I K <10% 100%** 50-75% 75-100% 10-25% 37°C
Sfi I B 75-100% 100% 75-100% 25-50% 75-100% 50°C
Sgf I (a) C 25-50% 25-50% 100% <10% <10% 37°C
Sin I A 100% 75-100% 50-75% 10-25% 100% 37°C
Sma I J <10% <10% 10% <10% 100% 25°C
SnaB I B 50-75% 100% 50-75% <10% 100% 37°C
Spe I B 75-100% 100% 75-100% 75-100% 100% 37°C
Sph I K 75-100% 75-100%** 100% 75-100% 10-25% 37°C
Ssp I E 10-25% 50-75% 50-75% 75-100% 50-75% 37°C
Stu I B 75-100% 100% 75-100% 50-75% 50-75% 37°C
Sty I F 25-50% 75-100% 75-100% 75-100% <10% 37°C
Taq I E 10-25% 25-50% 50-75% 50-75% 100% 65°C
Tru9 I F 75-100% 50-75% 75-100% 25-50% 25-50% 65°C

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Promega
Enzyme 		Buffer Supplied
with Enzyme 		4 CORE® Buffers MULTI-
CORE™ Buffer		 Enzyme Assay
Temperature
A B C D
Tth111 I B 50-75% 100% 75-100% 25-50% 100% 65°C
Vsp I D <10% 25-50% 75-100% 100% <10% 37°C
Xba I D 50-75% 75-100% 75-100% 100% 100% 37°C
Xho I D 25-50% 75-100% 75-100% 100% 10-25% 37°C
Xho II C 25-50% 25-50% 100% 10-25% <10% 37°C
Xma I B 50-75% 100% 25-50% <10% 50-75% 37°C
Xmn I B 75-100% 100% 75-100% 10-25% 75-100% 37°C

*Sac II exhibits 100% activity with linear substrates in buffer A. It exhibits very poor activity with supercoiled substrates in this buffer. 
**These buffers are not recommended due to star activity.

(a) U.S. Pat. No. 5,391,487 has been issued to Promega Corporation for Restriction Endonuclease Sgf I.

4-CORE is a trademark of Promega Corporation and is registered with the U.S. Patent and Trademark Office. MULTICORE is a trademark of Promega Corporation.

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